通过磁激活细胞分选(MACS)丰富 hiPSC 来源的心肌细胞。

Enriching Cardiomyocytes Derived from hiPSCs by Magnetic-Activated Cell Sorting (MACS).

机构信息

Departament of Medical Biotechnology, Faculty of Biochemistry, Biophysics and Biotechnology, Jagiellonian University, Krakow, Poland.

Jagiellonian University, Doctoral School of Exact and Natural Sciences, Krakow, Poland.

出版信息

Methods Mol Biol. 2024;2835:83-98. doi: 10.1007/978-1-0716-3995-5_8.

Abstract

Cardiomyocytes (CMs) derived from human-induced pluripotent stem cells (hiPSCs) are considered a promising platform for multiple applications, including disease modeling, regenerative medicine, screening of drug toxicity and investigation of cardiomyogenesis. Despite remarkable improvement in methodology enabling differentiation of hiPSCs into CMs, applied protocols generate heterogeneous cell populations composed of CMs along with differentiated non-cardiac cell-types and undifferentiated hiPSCs. Here we describea procedure of automated Magnetic-Activated Cell Sorting (autoMACS) for the purification of hiPSCs-derived CMs under sterile culture conditions. We illustrate that this approach led to a robust depletion of non-cardiac cells and enrichment of CMs, a result particularly crucial for hiPSC lines with poor cardiac differentiation efficiencies.

摘要

人心肌细胞(CMs)来源于人诱导多能干细胞(hiPSCs),被认为是一种有前途的平台,可用于多种应用,包括疾病建模、再生医学、药物毒性筛选和心肌发生研究。尽管在使 hiPSCs 分化为 CMs 的方法上取得了显著的进展,但应用的方案产生了异质的细胞群体,其中包括 CMs 以及分化的非心肌细胞类型和未分化的 hiPSCs。在这里,我们描述了一种在无菌培养条件下通过自动化磁激活细胞分选(autoMACS)对 hiPSCs 衍生的 CMs 进行纯化的程序。我们表明,这种方法导致非心肌细胞的大量耗竭和 CMs 的富集,对于心脏分化效率差的 hiPSC 系来说,这一结果尤为关键。

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